Dinitroxide biradical compounds as polarizing agents

ABSTRACT

The present invention relates to novel organic dinitroxide biradical compounds and their use as polarizing agents, in particular, in the techniques of Nuclear Magnetic Resonance (NMR) of solids or liquid samples and medical imaging.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/628,207, filed Jan. 2, 2020, which is a U.S. National Stage ofInternational Application No. PCT/EP2018,068002, filed Jul. 3, 2018,which claims priority under 35 U.S.C. § 119 to European PatentApplication No. 17179869.7, filed Jul. 5, 2017, the entire disclosuresof which are hereby incorporated by reference.

The present invention relates to novel organic dinitroxide biradicalcompounds and their use as polarizing agents, in particular, in thetechniques of Nuclear Magnetic Resonance (NMR) of solids or liquidsamples and medical imaging.

Nuclear magnetic resonance (NMR) is a very informative characterizationmethod that allows probing matter by detecting signals of nuclear spinsplaced in a sufficiently stable, homogeneous and intense magnetic field(up to tens of Tesla typically). This technique makes it possible torecord spectral data which, once interpreted, provide information at theatomic scale on the structure and the dynamics of the system studied.The use of NMR is widespread in chemistry, biology and materialsscience.

Several classes of NMR experiments have been developed over the years.Static solid-state NMR which stands for experiments conducted on staticsample holders, often filled with oriented samples. Magic Angle Spinning(MAS) solid-state NMR refers to experiments conducted on spinningsamples. This approach is based on the pneumatic rotation of the samplein order to induce a spatial averaging of anisotropic interactions. Theaxis of the rotation often corresponds, but not limited to, a preciseorientation with respect to the magnetic field called “magic angle”.Another class of experiments involve the detection of liquids and isbroadly referred to solution-state NMR. Finally, experiments aiming atperforming imaging are referred to Magnetic Resonance Imaging (MSI).

NMR detection offers a lot of information but suffers from a lack ofsensitivity in comparison with other analytical techniques. Theacquisition of signals of interest is therefore often long and/orrequires the use of important amount of material. This limitation isinherent to the properties of the nuclei, which are said to have a smallpolarization under standard experimental conditions. This lack ofsensitivity undoubtedly limits the use of NMR for the development of newmaterials (for catalysis, energy storage and conversion, etc.), as wellas its impact in biology and health related challenges.

To compensate for this lack of sensitivity, a technique ofhyperpolarization, called dynamic nuclear polarization (DNP), can beused. This phenomenon, discovered in 1953 on conductive materials usingweak magnetic fields, has gradually became a very active field ofresearch over the last decades, after it was shown that thishyperpolarization step could be combined with contemporary magneticfields (3 T-20 T and higher). The main two approaches are called“dissolution DNP” and “solid-state DNP”. The former polarizes the samplein the solid-state (often at very low temperature) and is followed by asample melting step, before NMR detection in the liquid state. The laterpolarizes the sample in the solid-state, and perform the NMR detectionin the solid-state as well, with static or spinning samples.

The development of solid-state DNP towards higher magnetic field, whilekeeping high resolution conditions (MAS), was pioneered in the group ofProf. Griffin at MIT. Thanks to major instrumental and theoreticaldevelopments, including access to high-power and high-frequencymicrowave sources, dynamic nuclear polarization applied to solid-statenuclear magnetic resonance appears as a promising solution forsignificantly increasing nuclear magnetization in many systems. Thisapproach makes it possible to record new type of data and thus, obtainstructural characteristics hitherto inaccessible. Currently, DNP (underMAS) measurements are typically performed at magnetic fields of about 1to 20 Tesla, and, in most cases, require measurements at low temperatureabout 100 K). The measurements are carried out in sample holders(rotors) with a capacity of 1-100 μ1.[1]

In high magnetic field DNP experiments, the systems of interest aregenerally dissolved, suspended or impregnated with a solution containingpolarizing agents. This solution is typically chosen as a function ofthe system to be studied, and for its specific characteristics in termsof DNP efficiency (quality of the glass formed at low temperature,relaxation time of nuclear and electronic spins). The polarizing agentmolecules contain paramagnetic centers with unpaired electrons, whichgive them an electron spin, about 660 times more polarized than anuclear proton spin. The application of a suitable microwave irradiationmakes it possible to transfer the magnetization of the electronic spinsto the surrounding nuclear spins and, thus, to significantly increasethe detected NMR signal.

Numerous polarizing agents have been developed over the years in orderto optimize the signal-to-noise increase obtained by DNP. Several DNPmechanisms under rotation at the magic angle and high magnetic fieldhave been described to date (K. R. Thurber and R. Tycko, “Theory forcross effect dynamic nuclear polarization under magic-angle spinning insolid state nuclear magnetic resonance: the importance of levelcrossings.,” J. Chem. Phys., vol. 137, no. 8, pp. 84508, Aug. 2012; F.Mentink-Vigier, U. Akbey, Y. Hovav, S. Vega, H. Oschkinat, and A.Feintuch, “Fast passage dynamic nuclear polarization on rotatingsolids,” J. Magn. Reson., vol. 224, pp. 13-21, Nov. 2012). Among thesedifferent mechanisms, “Cross-Effect” (CE) combined with the use ofbiradicals as polarizing agents is currently the most promising in termsof sensitivity.

The modalities for the transfer of the polarization of the electronstowards the nuclei depend on the nature of the paramagnetic centersused. The objective is to maximize the polarization transfer whileminimizing the time for such transfer under general experimentalconditions: in the presence of a strong magnetic field, and/or rapidrotation of the sample. There are several magnetization transfermechanisms under MAS, such as “Solid-Effect” (SE). “Cross-Effect” (CE)and “Overhauser Effect” (OE).

Under MAS-DNP experimental conditions as of today, which correspond to amagnetic field of about 5 to 20 T (or more), a sample rotation in the1-40 kHz frequency range (or more), and a temperature of about 100 K,the CE is currently the method that offers the best results for DNPexperiments under MAS. This type of experiment usually involves the useof polarizing agents that typically contain two paramagnetic centers,for example, two nitroxide entities bound by a bridge. This bridge,which ensures a substantial interaction between the electronic spins, isnecessary for the CE mechanism. This interaction (dipole or/-exchangeinteraction) must be significant but not exceed the Larmor frequency ofthe targeted nucleus, as too much coupling leads to a loss of efficiencyof the mechanism.

The distance between the electronic spins, and therefore the coupling,is not the only important criterion for optimizing the polarizationgain. Like any paramagnetic center the magnetic properties of anunpaired spin are characterized by a g-tensor which relates to themolecular structure. In order for the CE to be active, it is alsonecessary for the planes defined by the nitroxide entities not to beparallel, in other words that the g-tensors are not collinear. Theelectron spin relaxation times (T_(1e)/T_(2e)) are also importantparameters, as well as the nuclear spin density (e.g. protons), thehyperfine couplings and the associated relaxation times (in theabsence/presence of paramagnetic doping). Ultimately, it is a complexset of parameters that governs the effectiveness of a DNP experiment.

The molecules most commonly used correspond to bi-nitroxide biradicals(TOTAPOL, the bTbK family, the bTUrea family) or mixed radicals(Tempo-Trityl and Tempo-BDPA). One of the first examples of binitroxidesfor DNP applications was:

-   -   1-(TEMPO-4-oxyl)-3-(TEMPO-4-amino)-propan-2-ol (TOTAPOL)

which is compatible with experiments in aqueous media, including saltsolutions commonly used in the study of proteins and nucleic acids. (C.Song, K.-N. N. Hu, C.-G. Joo, T. M. Swager, R. G. Griffin, J. Am. Chem.Soc, vol. 128, no. 35, pp. 11385-90, September 2006), and the bTUrea(bis-TEMPO-Urea) family

(C. Sauvee, G. Casano, S. Abel, A. Rockenbauer, D. Akhmetzyanov, H.Karoui, D. Ski, F. Aussenac, W. Maas, R. T. Weber, T. Prisner, M. Rosay,P. Tordo, 0. Ouari, Chem.—A Eur. J., vol. 22, no. 16, pp. 5598-5606,April 2016), in which link units connecting the nitroxide units are anethylene glycol unit and a urea bridge, respectively. Note that the useof such bridge in bTUrea has reduced the distance between the electronicspins and rigidified the molecule i.e. the relative orientation of thenitroxide plans, as compared to the TOTAPOL case. Consequently, theinteraction between the nitroxides is stronger and their relativeorientation better defined, contributing to an increase of the DNPefficiency, which is consistent with the simulations recently reported(F. Mentink-Vigier, U. Akbey, H. Oschkinat, S. Vega, A. Feintuch, J.Magn. Reson., vol. 258, pp. 102-120, September 2015).

In parallel, another family of polarizing agents was introduced: thebTbK family, for which the linking motif is a ketal bridge.

This ketal bridge imparts a great rigidity to the molecule and fixesalmost totally the relative orientation of the two nitroxides and theirassociated g tensors. It is of note that this relative orientation isconsidered in the literature as close to the ideal solution. (Y.Matsuki, T. Maly, O. Ouari, H. Karoui, F. Le Moigne, E. Rizzato, S.Lyubenova, J. Herzfeld, T. F. Prisner, P. Tordo, R. G. Griffin, Angew.Chem. Int. Ed. Engl., vol. 48, no. 27, pp. 4996-5000, January 2009).

At this stage, it is important to note that the polarizing agentsdeveloped up to now are evaluated by measuring the ratio of the signalwith and without microwave irradiation as well as the rise time inpolarization.

The polarizing agents proposed up to now offer modest performances whenthe magnetic field is >9 T and/or the MAS frequency is >10 kHz. Theirperformance drops significantly at higher magnetic field, due to lowerpolarization gain and slower polarization speed (at higher magneticfield).

It is of high interest to develop new polarizing agents with new typesof chemical bridges that will enhance the polarization transfer whileminimizing the time for such transfer. Therefore, there is a real needfor new polarizing agents having a chemical bridge that combines both astrong interaction between the electronic spins, a good relativeorientation, interesting relaxation properties while maintaining goodsolubility in a DNP-compatible solvent and compatible with the intendedapplication.

The present invention addresses these needs among others by providingcompounds of formula (I)

wherein

-   -   X₁ is O, C(R⁶R⁷), NR⁸, S;    -   X₂ is O, SO₂, or —NR⁹, CH₂;    -   X₃ is C or N with the proviso that when X₃ is N, then R⁵ is not        present in the molecule;    -   R⁵, R⁶, R⁷, R⁸ and R⁹ are, independently, H; a substituted or        unsubstituted, linear, branched or cyclic C₁₋₆ aliphatic group;        —(CH₂)_(n)—COOH with n being an integer from 1 to 10, —OH, —NH₂,        —N₃, —C≡CH, P(O)(OH)₂, P(O)(OR¹¹)₂, P(O)R¹¹ ₂, —SSO₂Me,        —(CH₂—CH₂—O)_(m) —CH₃ or —(CH₂—CH₂—O)_(m)—H with m being an        integer from 1 to 500, preferably from 1 to 100, more preferably        from 1 to 10, or

-   -   with p being an integer from 0 to 7 and q an integer from 1 to        500;    -   Q₁ is a cyclic or acyclic nitroxide radical, as represented        below

-   -   R¹ and R² are, independently, a substituted or unsubstituted,        linear, branched or cyclic C₁₋₆ aliphatic group; a substituted        or unsubstituted linear, branched or cyclic hetero-aliphatic        group comprising 5 carbon atoms and one heteroatom or        heteroatomic group, respectively, selected from O, S, —NR¹⁰—,        P(O)(OR¹¹) and P(O)(R¹¹) substituted or unsubstituted aryl;        substituted or unsubstituted heteroaryl, or substituted or        unsubstituted 2,2,7,7-tetramethyl isoindolinoxyl, or substituted        or unsubstituted 2,2,7,7-tetraethyl isoindolinoxyl; or    -   R¹ and R² are joined, as indicated by

to form together with the nitrogen atom to which they are bound a 5- to8-membered heterocyclic ring and which may contain an additionalheteroatom or heteroatomic group

-   -   selected from P(O)(OR¹¹), P(O)(R¹¹), O, S, N⁺—O⁻, NH, N(C₁₋C₆        alkyl) wherein the alkyl is straight, branched or cyclic,        wherein the heterocyclic ring bears from one substituent to the        maximum number of substituent on the carbon atoms and optionally        contains one double bond; and    -   with the proviso that the two groups R¹ or R² together do not        contain more than one hydrogen alpha to the (N—O) group;    -   R¹⁰ is hydrogen, hydroxyl; substituted or unsubstituted linear,        branched or cyclic C₁₋₆ alkyl; C₁₋₆ alkylcarbonyl; substituted        or unsubstituted aryl sulfinyl; or substituted or unsubstituted        aryl sulfonyl;    -   R¹¹ is linear or branched C₁₋₁₈ alkyl, H or an alkali metal; and    -   wherein the point of attachment of the X₂ atom by a single bond,        as indicated by

is to a primary or secondary non-olefmic or aromatic carbon atom ofeither R¹ or R², or to a carbon atom of the 5- to 8-memberedheterocyclic ring formed by the joining of R¹ and R²;

-   -   R³ or R⁴ is linked to the double bond C═X₃ in the compound of        formula (I) as represented above, wherein R³ and R⁴ are,        independently, a substituted or unsubstituted, linear, branched        or cyclic C₁₋ ₆ aliphatic group; a substituted or unsubstituted        linear, branched or cyclic hetero-aliphatic group comprising 5        carbon atoms and one heteroatom or heteroatomic group,        respectively, selected from O, S, —NR¹⁰—, P(O)(OR¹¹) and        P(O)(R¹¹); substituted or unsubstituted aryl; substituted or        unsubstituted heteroaryl, or substituted or unsubstituted 2,2,7        ,7-tetramethyl isoindolinoxyl, substituted or unsubstituted        2,2,7,7-tetraethyl isoindolinoxyl; or    -   R³ and R⁴ are joined, through the double bond C═X₃ to form        together with the nitrogen atom to which they are bound a 5- to        8-membered heterocyclic ring and which may contain an additional        heteroatom or heteroatomic group selected from P(O)(OR¹¹),        P(O)(R¹¹), O, S, N⁺—O^(—), NH, N(C₁₋C₆ alkyl) wherein the alkyl        is straight, branched or cyclic, wherein the heterocyclic ring        bears from one substituent to the maximum number of substituent        on the carbon atoms and optionally contains one double bond; and    -   with the proviso that the two groups R³ or R⁴ together do not        contain more than one hydrogen alpha to the (N—O) group;    -   with the proviso that a compound of formula

is excluded.

By this invention, the inventors have designed a new family of powerfulpolarizing agents with high magnetic field and high frequency MAS. Thisnew generation of molecules, based on symmetrical or asymmetricdinitroxides, uses in particular a new type of chemical bridge.

The chemical bridge or linkage presented in the compounds of formula (I)is a conjugate bridge that provides stiffness and rigidity which reducesthe distance between the nitroxide units and promotes the efficienttransfer of polarization. It is worth noting that the optimal polarizingagent structure is a complex multi-parameter problem which implies forinstance, the distance between the two nitroxides, the intensity of theJ-exchange interaction, the relative orientation between the twonitroxides, as well as the associated electronic relaxation times anddeuteration level of the polarizing agent.

The design of these new compounds did not consist solely in trying tomaximize the ^(ε)on/off amplification factor but also to minimizedepolarization effects while maximizing the effectiveness of thetransfer of polarization. The inventors aimed at maximizing thesensitivity under DNP conditions, defined by ϵ_(B)/√{square root over(T_(B) ^(T))}⁺ where ε_(B) and T_(B) represent the real enhancementfactor corrected from depolarization effect and the polarization builduptime respectively. (F. Mentink-Vigier, S. Paul, D. Lee, A. Feintuch, S.Hediger, S. Vega and G. De Paepe, Phys. Chem. Chem. Phys. , 2015, 17,21824. and F. Mentink-Vigier, S. Vega and G. De Paepe, Phys. Chem. Chem.Phys., 2017, 19, 3506-3522.)

With the goal to increase the interaction between unpaired electrons(called dipolar coupling and/or J exchange interaction) and constrainthe relative orientation of the two paramagnetic centers (e.g. theplanes of the TEMPO groups), the inventors found that a conjugatedchemical bridge as in compounds of formula (I), reduces the number ofatoms involved in the bridge. Such conjugated bridge brings the twoelectron spins closer while maintaining a suitable relative orientationbetween the two nitroxide groups (N—O.) (or their correspondingg-tensors) to perform CE DNP.

Due to the strong interaction between the resulting electronic spins andtheir relative orientation, the polarization transfer efficiency definedby ϵ_(B) (and not the amplification factor ϵon/off) can be maximized andthe polarization rise time T_(B) can be minimized.

The term “C₁₋₆ aliphatic group”, as used herein, includes both saturatedand unsaturated, non aromatic, straight chain (i.e. unbranched),branched, acyclic and cyclic (i.e. carbocyclic) hydrocarbons having 1-6carbon atoms, which are optionally substituted with one or morefunctional groups. The term “aliphatic”, as used herein, includes C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₇ cycloalkyl, C₃₋₇ cycloalkenyland C₅₋₇ cycloalkynyl moieties. Thus, as used herein, the term “C₁₋₆alkyl” includes straight, branched and cyclic hydrocarbons having 1-6carbon atoms. Similarly, the terms “C₂₋₆ alkenyl” and “C₂₋₆ alkynyl”,includes straight, branched, unsaturated and cyclic hydrocarbons having2-6 carbon atoms and at least one unsaturation (a double and/or triplebond).

Substituents in a substituted C₁₋₆ aliphatic group include C₁₋₄hydroxyalkyl, C₁₋₄ alkoxy, C₁₋₄ alkylthio; C₁₋₄ alkylcarbonyl; C₁₋₄alkylsulfonyl; C₁₋₄ alkylsulfinyl, whose alkyl moieties may be partiallyor completely halogenated with independent halogen atoms; halogen;hydroxyl; thiol; nitro; cyano; —NR¹⁰ ₂; ═NR¹⁰; —COOH; COOR¹⁰; —CONR¹⁰ ₂and SO₂NR¹⁰ ₂, wherein R¹⁰ is independently hydrogen, hydroxyl,substituted or unsubstituted linear, branched or cyclic C₁₋₆ alkyl, C₁₋₆alkylcarbonyl, substituted or unsubstituted aryl sulfinyl, orsubstituted or unsubstituted aryl sulfonyl. The number of substituentspresent in a substituted C₁₋₆ aliphatic group may be up to the number ofthe hydrogen atoms available for a susbtitution, but is preferably 1-2,more preferably 1.

The term “C₁₋₆ alkyl” as used herein, refers to saturated, straight- orbranched-chain or cyclic hydrocarbon radicals derived from a hydrocarbonmoiety containing from 1 to 6 carbon atoms by removal of a singlehydrogen atom. Examples of C₁₋₆ alkyl radicals include methyl, ethyl,n-propyl, isopropyl, n-butyl, iso-butyl, sec-butyl, sec-pentyl,isopentyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, sec-hexyl,cyclopropyl, cyclopropylmethyl, cyclopentyl and cyclohexyl. The term“C₁₋₄ alkyl”, as used herein, refers to saturated, straight- orbranched-chain hydrocarbon radicals derived from a hydrocarbon moietycontaining from 1 to 4 carbon atoms by removal of a single hydrogenatom. Examples of C₁₋₄ alkyl radicals include methyl, ethyl, n-propyl,isopropyl, n-butyl, iso-butyl, sec-butyl. The term “C₁₋₁₈ alkyl” as usedherein, refers to saturated, straight- or branched-chain hydrocarbonradicals derived from a hydrocarbon moiety containing from 1 to 18carbon atoms by removal of a single hydrogen atom.

The term “C₃₋₇ cycloalkyl” refers to cyclic hydrocarbon radicals derivedfrom a hydrocarbon moiety containing from 3 to 7 carbon atoms by removalof a single hydrogen atom. Examples of C₃₋₇ cycloalkyl includecyclopropyl, cyclopropylmethyl, cyclobutyl, cyclobutylmethyl,cyclopentyl, cyclopentylmethyl, cyclohexyl, and cyclohexylmethyl.

The above-mentioned alkyl or cycloalkyl groups may bear one or moresubstituents. Substituents in a substituted C₁₋₆ alkyl, C₁₋₄ alkyl orC₃₋₇ cycloalkyl group include C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₁₋₄alkylthio; C₁₋₄ alkylcarbonyl; C₁₋₄ alkylsulfonyl; C₁₋₄ alkylsulfinyl,whose alkyl moieties may be partially or completely halogenated withindependent halogen atoms; halogen; hydroxyl; thiol; nitro; cyano; —NR¹⁰₂; ═NR¹⁰; —COOH; COOR¹⁰; —CONR¹⁰ ₂ and SO₂NR¹⁰ ₂, wherein R¹⁰ isindependently hydrogen, hydroxyl, substituted or unsubstituted linear,branched or cyclic C₁₋₆ alkyl, C₁₋₆ alkylcarbonyl, substituted orunsubstituted aryl sulfinyl, or substituted or unsubstituted arylsulfonyl. The number of substituents present in a substituted C₁₋₆ alkylgroup may be up to the number of the hydrogen atoms available for asusbtitution, but is preferably 1-2, more preferably 1.

The term “C₂₋₆ alkenyl”, as used herein, refers to a monvalent groupderived from a straight- or branched-chain or cyclic hydrocarbon moietyhaving at least one carbon-carbon double bond. The C₂₋₆ alkenyl contains2 to 6 carbon atoms. C₂₋₆ alkenyl groups include, for example, ethenyl,propenyl, isopropenyl, 1- or 2-butenyl, 1-methyl-2-buten-1-yl andcyclopentenyl. C₂₋₄ alkenyl, as used herein, refers to a monvalent groupderived from a straight- or branched-chain or cyclic hydrocarbon moietyhaving at least one carbon-carbon double bond. The C₂₋₄ alkenyl contains2 to 4 carbon atoms. C₂₋₄ alkenyl groups include, for example, ethenyl,propenyl, isopropenyl, 1- or 2-butenyl.

The above-mentioned C₂₋₆ or C₂₋₄ alkenyl groups may bear one or moresubstituents. Substituents in a substituted C₂₋₆ or C₂₋₄ alkenyl groupinclude C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₁₋₄ alkylthio; C₁₋₄alkylcarbonyl; C₁₋₄ alkylsulfonyl; C₁₋₄ alkylsulfinyl, whose alkylmoieties may be partially or completely halogenated with independenthalogen atoms; halogen; hydroxyl; thiol; nitro; cyano; —NR¹⁰ ₂; ═NR¹⁰;—COOH; COOR¹⁰; —CONR¹⁰ ₂ and SO₂NR¹⁰ ₂, wherein R¹⁰ is independentlyhydrogen, hydroxyl, substituted or unsubstituted linear, branched orcyclic C₁₋₆ alkyl, C₁₋₆ alkylcarbonyl, substituted or unsubstituted arylsulfinyl, or substituted or unsubstituted aryl sulfonyl. The number ofsubstituents present in a substituted C₂₋₆ or C₂₋₄ alkenyl group may beup to the number of the hydrogen atoms available for a susbtitution, butis preferably 1-2, more preferably 1.

The term “C₂₋₆ alkynyl”, as used herein, refers to a monvalent groupderived from a straight- or branched-chain or cyclic hydrocarbon moietyhaving at least one carbon-carbon triple bond. The C₂₋₆ alkynyl contains2 to 6 carbon atoms. C₂₋₆ alkynyl groups include, for example, ethynyl,2-propynyl (propargyl), 1-propynyl and cyclohexynyl. C₂₋₄ alkynyl, asused herein, refers to a monvalent group derived from a straight- orbranched-chain or cyclic hydrocarbon moiety having at least onecarbon-carbon triple bond. The C₂₋₄ alkynyl groups contain 2 to 4 carbonatoms. C₂₋₄ alkynyl groups include, for example, ethynyl, 2-propynyl(propargyl), 1-propynyl.

The above-mentioned C₂₋₆ or C₂₋₄ alkynyl groups may bear one or moresubstituents. Substituents in a substituted C₂₋₆ or C₂₋₄ alkynyl groupinclude C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₁₋₄ alkylthio; C₁₋₄alkylcarbonyl; C₁₋₄ alkylsulfonyl; C₁₋₄ alkylsulfinyl, whose alkylmoieties may be partially or completely halogenated with independenthalogen atoms; halogen; hydroxyl; thiol; nitro; cyano; —NR¹⁰ ₂, ═NR¹⁰,—COOH, COOR¹⁰, —CONR¹⁰ ₂ and SO₂NR¹⁰ ₂, wherein R¹⁰ is independentlyhydrogen, hydroxyl, substituted or unsubstituted linear, branched orcyclic C₁₋₆ alkyl, C₁₋₆ alkylcarbonyl, substituted or unsubstituted arylsulfinyl, or substituted or unsubstituted aryl sulfonyl. The number ofsubstituents present in a substituted C₂₋₆ or C₂₋₄ alkynyl group may beup to the number of the hydrogen atoms available for a susbtitution, butis preferably 1-2, more preferably 1.

The term “C₁₋₆ heteroaliphatic group”, as used herein, includes bothsaturated and unsaturated, non aromatic, straight chain (i.e.unbranched), branched, acyclic and cyclic (i.e. heterocyclic)hydrocarbons, which are optionally substituted with one or morefunctional groups and that contain one oxygen, sulfur, substitutednitrogen or substituted phosphorous atom in place of one carbon atom. Aheteroaliphatic group according to the invention has 1-5 carbon atoms.The term “C₁₋₆ heteroaliphatic group” includes heteroalkyl,heteroalkenyl, heteroalkynyl and heterocyclyl moieties. Thus, as usedherein, the term “heteroalkyl” includes straight, branched and cyclicalkyl groups as defined herein, which are substituted with one or morefunctional groups, and that contain one oxygen, sulfur, nitrogen orphosphorous atom in place of one carbon atom. An analogous conventionapplies to other generic terms such as “heteroalkenyl” and“heteroalkynyl”. The substituent of the nitrogen atom is the substituentR^(a), as defined in the context of the present invention. Phosphorousis present as P(O)OH, P(O)(C₁₋₁₈ alkoxy) or P(O)(C₁₋₁₈ alkyl). The term“substituted C₁₋₆ heteroaliphatic group” denotes that one or more carbonatoms may also bear a substituent.

Substituents on carbon atoms of the C₁₋₆ heteroaliphatic group includeC₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₁₋₄ alkylthio; C₁₋₄ alkylcarbonyl; C₁₋₄alkylsulfonyl; C₁₋₄ alkylsulfinyl, whose alkyl moieties may be partiallyor completely halogenated with independent halogen atoms; halogen;hydroxyl; thiol; nitro; cyano; —NR¹⁰ ₂, ═NR¹⁰, —COOH, COOR¹⁰, —CONR¹⁰ ₂and SO₂NR¹⁰ ₂, wherein R¹⁰ is independently hydrogen, hydroxyl,substituted or unsubstituted linear, branched or cyclic C₁₋₆ alkyl, C₁₋₆alkylcarbonyl, substituted or unsubstituted aryl sulfinyl, orsubstituted or unsubstituted aryl sulfonyl. The number of substituentspresent in a substituted C₂₋₆ or C₂₋₄ alkynyl group may be up to thenumber of the hydrogen atoms available for a susbtitution, but ispreferably 1-2, more preferably 1.

The term “aryl”, as used herein, refers to stable mono- or bicyclic ringsystem having 5-10 ring atoms, of which all the ring atoms are carbon,and which may be substituted or unsubstituted. Aryl includes, forexample, phenyl, biphenyl and naphtyl, which may bear one or moresubstituents;

Aryl substituents include C₁₋₆ alkyl, C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy,C₁₋₄ alkylthio; C₁₋₄ alkylcarbonyl; C₁₋₄ alkylsulfonyl; C₁₋₄alkylsulfinyl, whose alkyl moieties may be partially or completelyhalogenated with independent halogen atoms; halogen; hydroxyl; thiol;nitro; cyano; —NR¹⁰ ₂; ═NR¹⁰; —COOH; COOR¹⁰; —CONR¹⁰ ₂ and SO₂NR¹⁰ ₂ ,wherein R¹⁰ is independently hydrogen, hydroxyl, substituted orunsubstituted linear, branched or cyclic C₁₋₆ alkyl, C₁₋₆ alkylcarbonyl,substituted or unsubstituted aryl sulfinyl, or substituted orunsubstituted aryl sulfonyl.

2,2,7,7-Tetramethyl isoindolinoxyl and 2,2,7,7-tetraethyl isoindolinoxylsubstituents on the benzene ring are the same as the aryl substituents.

Preferably aryl is unsubstituent phenyl or phenyl substituted with oneof the above substituents.

The term “heteroaryl”, as used herein, refers to stable mono- orbicyclic ring system having 5-12 ring atoms, of which one ring atom isselected from S, O, and N and the remaining atoms are carbon. 0, 1 or 2ring atoms are additional heteroatoms independently selected from S, O,and N and the remaining atoms are carbon. The heteroaryl radical may bejoined to the rest of the molecule via any of the ring atoms. Exemplaryheteroaryls include pyrrolyl, pyrazolyl, imidazolyl, pyridinyl,pyrimidinyl, pyrazinyl, pyridazinyl, triazinyl, tetrazinyl,pyrrolizinyl, indolyl, quinolinyl, isoquinolinyl, benzoimidazolyl,indazolyl, quinolinyl, isoquinolinyl, quinolizinyl, cinnolinyl,quinazolynyl, phtalazinyl, naphthridinyl, quinoxalinyl, thiophenyl,thianaphthznyl, furanyl, benzofuranyl, benzothiazolyl, thiazolynyl,isothiazolyl, thiadiazolynyl, oxazolyl, isoxazolyl, oxadiazolyl andoxadiazolyl may bear one or more substituents. Heteroaryl substituentsinclude C₁₋₆ alkyl, C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₁₋₄ alkylthio; C₁₋₄alkylcarbonyl; C₁₋₄ alkylsulfonyl; C₁₋₄ alkylsulfinyl, whose alkylmoieties may be partially or completely halogenated with independenthalogen atoms; halogen; hydroxyl; thiol; nitro; cyano; —NR¹⁰ ₂ ; ═NR¹⁰;—COOH; COOR¹⁰; —CONR¹⁰ ₂ and SO₂NR¹⁰ ₂ , wherein R¹⁰ is independentlyhydrogen, hydroxyl, substituted or unsubstituted linear, branched orcyclic C₁₋₆ alkyl, C₁₋₆ alkylcarbonyl, substituted or unsubstituted arylsulfinyl, or substituted or unsubstituted aryl sulfonyl.

Preferably heteroaryl is pyrrol, pyrazolyl, imidazolyl, pyridinyl,pyrimidinyl, pyrazinyl, pyridazinyl, furanyl and thiophenyl, eachsubstituted or unsubstituted with one of the above substituents.

The term “heterocyclic ring”, as used herein, refers to a non aromatic,partially unsaturated or fully saturated, 5- to 8-membered ring. Theseheterocyclic rings include those having 1 to 3 heteroatoms independentlyselected from oxygen, sulfur, phosphorous and nitrogen, in which thenitrogen, phosphorous and sulfur heteroatoms may optionally be oxidizedand the nitrogen heteroatom may optionally be quatemized. In certainembodiments, the term heterocyclic ring refers to a non aromatic 5-, 6-,7- or 8-membered ring wherein at least one ring atom is a heteroatomselected from oxygen, sulfur, phosphorous and nitrogen wherein thesubstituent of the nitrogen atom is R¹⁰ as defined above, andphosphorous is present as P(O)OH, P(O)OR¹¹ or P(O)R¹¹, with R¹¹ being alinear or branched C₁₋₁₈ alkyl. The remaining atoms of the heterocyclicring are carbon atoms, and said ring is joined to the rest of themolecule via any of the carbon ring atoms.

Substituents on carbon atoms of the heterocyclic rings include C₁₋₆alkyl, C₁₋₄ hydroxyalkyl, C₁₋₄ alkoxy, C₁₋₄ alkylthio; C₁₋₄alkylcarbonyl; C₁₋₄ alkylsulfonyl; C₁₋₄ alkylsulfinyl, whose alkylmoieties may be partially or completely halogenated with independenthalogen atoms; halogen; hydroxyl; thiol; nitro; cyano; —NR¹⁰ ₂ ; ═NR¹⁰;—COOH; COOR¹⁰; —CONR¹⁰ ₂ and SO₂NR¹⁰ ₂ , wherein R¹⁰ is independentlyhydrogen, hydroxyl, substituted or unsubstituted linear, branched orcyclic C₁₋₆ alkyl, C₁₋₆ alkylcarbonyl, substituted or unsubstituted arylsulfinyl, or substituted or unsubstituted aryl sulfonyl. Furthermore,any two geminal substituents of heterocyclic rings may be joined to formtogether with the secondary carbon atom to which they are bound acyclopentane or cyclohexane ring which may be further substituted withone or more substituents as defined above for alkyl groups, or a 5- or6-membered heterocyclic ring comprising one atom or group selected fromO, S, NR¹⁰ with R¹⁰ as previously defined, P(O)OH, P(O)(C₁₋₆ alkyl) andP(O)(OC₁₋₆ alkyl) which may be further substituted on one or more carbonatoms with a substituent as defined above in this paragraph.

The term “hydroxy” or “hydroxyl”, as used herein, refers to —OH. Theterm “C₁₋₄ alkoxy” refers to a group of formula —OR^(a) wherein R^(a) isC₁₋₄ alkyl. The term “C₁₋₁₈ alkoxy” refers to a group of formula —OR^(b)wherein R^(b) is C₁₋₁₈ alkyl.

The term “C₁₋₄ hydroxyalkyl” refers to a C₁₋₄ alkyl group bearing onehydroxyl in place of any hydrogen atom.

The term “cyano”, as used herein, refers to —CN. The term “direct bond”or “bond” refers to a single, double or triple bond between two groups.In certain embodiments, a direct bond refers to a single bond betweentwo groups.

The terms “halo” and “halogen”, as used herein, refer to an atomselected from fluorine (fluoro —F), chlorine (chloro —Cl), bromine(bromo —Br), and (iodine (iodo —I).

The term “nitro”, as used herein, refers to —NO₂.

The term “nitroxide”, as used herein, refers to a stable cyclic oracyclic compound comprising at least one aminoxyl group. In certainembodiments, a stable nitroxide refers to a chemically stable nitroxidewhich may be obtained in pure form, stored and handled in thelaboratory. In certain embodiments, a stable nitroxide refers to acyclic or acyclic nitroxide which contains two groups which do notcontain alpha hydrogens. Exemplary cyclic or acyclic nitroxides areprovided, for exemple, in J. F. W. Keana, Chemical Reviews, 1978, 78,pp. 37-64.

A “dinitroxide biradical ”or a “dintroxide compound”, as used herein,refers to a stable cyclic or acyclic compound comprising two aminoxylgroups in two separate nitroxide containing moieties.

The term “C₁₋₄ alkylsulfinyl” and “C₁₋₆ alkylsulfinyl”, as used herein,refers to a group of formula C₁₋₄ alkyl-S(═O)— and C₁₋₆ alkyl-S(═O)—,respectively.

The term “aryl sulfinyl”, as used herein, refers to aryl-S(═O)—.

The term “C₁₋₄ alkyl sulfonyl” and “C₁₋₆ alkyl sulfonyl”, as usedherein, refers to a group of formula C₁₋₄ alkyl-S(═O)₂— and C₁₋₆alkyl-S(═O)₂—, respectively.

The term “aryl sulfonyl”, as used herein, refers to aryl-S(═O)₂—.

In a first embodiment of the invention, in the compounds of formula (I),Q₁ is a cyclic nitroxide radical, as represented below

wherein

R¹ and R² are joined, as indicated by

to form together with the nitrogen atom to which they are bound a 5- to8-membered heterocyclic ring and which may contain an additionalheteroatom or heteroatomic group selected from P(O)(OR¹¹), P(O)(R¹¹), O,S, N⁺−O⁻, NH, N(C₁₋C₆ alkyl) wherein the alkyl is straight, branched orcyclic, wherein the heterocyclic ring bears from one substituent to themaximum number of substituent on the carbon atoms and optionallycontains one double bond; and

-   -   with the proviso that the two groups R¹ or R² together do not        contain more than one hydrogen alpha to the (N—O) group; and    -   R¹¹ is linear or branched C₁₋₁₈ alkyl, H or an alkali metal.

In this first embodiment, X₁, X₂, X₃, R³ -R¹⁰, m, n, p and q are asdescribed above.

In this first embodiment, Q₁ is a nitroxide-containing a 5- to8-membered heterocyclic ring, substituted at least at all positionsalpha to the (N—O) group, selected from

wherein

-   -   z is 1 or 2;    -   Y is selected from —O—, —S—, —NR with R being hydrogen,        substituted or unsubstituted linear, branched or cyclic C₁₋₆        alkyl, or —CHR—with R being hydrogen, substituted or        unsubstituted linear, branched or cyclic C₁₋₆ alkyl; ═C    -   R¹³ and R¹⁴ are, independently, hydrogen, hydroxyl, substituted        or unsubstituted linear, branched or cyclic C₁₋₆ alkyl;    -   R¹² is, independently, hydrogen, hydroxyl, substituted or        unsubstituted linear, branched or cyclic C₁₋₆ alkyl, or    -   two geminal R¹² are joined to form together with the secondary        carbon to which they are bound a substituted or unsubstituted        cyclopentane, cyclohexane, a 5- or 6-membered heterocyclic ring        wherein at least one ring atom is oxygen.

In this first embodiment, Q₁ is, preferably,

wherein M is an alkali metal selected in the group consisting of lithium(Li), sodium (Na), potassium (K), rubidium (Rb) and cesium (Cs).Preferably, M is lithium (Li), sodium (Na) and potassium (K).

In a second embodiment of the invention, in the compounds of formula(I), Q₁ is an acyclic nitroxide radical, as represented below

wherein

R¹ and R² are, independently, a substituted or unsubstituted, linear,branched or cyclic C₁₋₆ aliphatic group; a substituted or unsubstitutedlinear, branched or cyclic hetero-aliphatic group comprising 5 carbonatoms and one heteroatom or heteroatomic group, respectively, selectedfrom O, S, —NR¹⁰—, P(O)(OR¹¹) and P(O)(R¹¹); substituted orunsubstituted aryl; substituted or unsubstituted heteroaryl, orsubstituted or unsubstituted 2,2,7,7-tetramethyl isoindolinoxyl; orsubstituted or unsubstituted 2,2,7,7-tetraethyl isoindolinoxyl;

with the proviso that the two groups R¹ or R² together do not containmore than one hydrogen alpha to the (N—O.) group.

In this second embodiment, X₁, X₂, X₃, R³-R¹¹, m, n, p and q are asdescribed above. In this second embodiment, Q₁ may be an acyclicnitroxide radical wherein R¹ and R² can be aliphatic, heteroaliphatic,aromatic, heteroaromatic groups wherein “aliphatic” is a substituted orunsubstituted linear, branched or cyclic C₁₋C₆ aliphatic group; and“heteroaliphatic” is a substituted or unsubstituted linear, branched orcyclic group heteroaliphatic group comprising 5 C atoms and oneheteroatom or heterogroup, respectively, selected from O, S, N(R¹²),P(R¹²), P(═O)OH and P(═O)O(C₁₋₆ alkyl); “aromatic” and “heteroaromatic”are substituted or unsubstituted “aryl” and “heteroaryl” respectively,as defined above; with the proviso that the two groups, which areselected from heteroaliphatic, aromatic or heteroaromatic group and areattached to the nitrogen atom of the (N—O.) group, together do notcontain more than one hydrogen alpha to the (N—O.) group; and anitroxide-containing 5- to 8-membered heterocyclic ring, which maycontain an additional heteroatom or heteroatomic group selected from O,S, N⁺—O⁻, NH, N(C₁₋₆ alkyl) wherein the alkyl is straight, branched orcyclic, and may contain one double bond and is substituted at least atall positions alpha to the (N—O) group, the substituents being selectedfrom those mentioned above in the context of the definition of“heterocyclic group”.

In this second embodiment, Q₁ is, preferably,

wherein

“Ar” is substituted or unsubstituted aryl; “Al” is a substituted orunsubstituted linear, branched or cyclic C₁₋₆ aliphatic group; and “Het”is a substituted or unsubstituted heteroaryl as defined herein.

Examples of Q₁ in this second embodiment are:

In a third embodiment of the invention, R³ and R⁴ are joined, throughthe double bond C═X₃ to form together with the nitrogen atom to whichthey are bound a 5- to 8-membered heterocyclic ring and which maycontain an additional heteroatom or heteroatomic group selected fromP(O)(OR¹¹), P(O)(R¹¹), O, S, N⁺—O⁻, NH, N(C₁₋C₆ alkyl) wherein the alkylis straight, branched or cyclic, wherein the heterocyclic ring bearsfrom one substituent to the maximum number of substituent on the carbonatoms and optionally contains one double bond; and

with the proviso that the two groups R³ or R⁴ together do not containmore than one hydrogen alpha to the (N—O.) group.

In this third embodiment, X₁, X₂, X₃, R⁵-R¹⁰, m, n, p and q are asdescribed above.

In this third embodiment, R³ and R⁴ are joined, through the double bondC═X₃ to form together with the nitrogen atom to which they are bound a5- to 8-membered heterocyclic ring, as represented below

wherein

-   -   z is 1 or 2;    -   X₃ is C or N with the proviso that when X₃ is N, then R⁵ is not        present in the molecule;    -   R⁵ is H; (CH₂ )_(n)—COOH with n being an integer from 1 to 10,        —OH, —NH₂, —(CH₂—CH₂—O)_(m)—CH₃ or (CH₂—CH₂—O)_(m)—H with m        being an integer from 1 to 10;    -   R¹⁵ and R¹⁶ are, independently, hydroxyl, substituted or        unsubstituted linear, branched or cyclic C₁₋₆ alkyl; or    -   two geminal R¹⁵ or R¹⁶ are joined to form together with the        secondary carbon to which they are bound a substituted or        unsubstituted cyclopentane, cyclohexane, a 5- or 6-membered        heterocyclic ring wherein at least one ring atom is oxygen.

In this third embodiment, preferably R³ and R⁴ are joined and formtogether with the nitrogen atom to which they are bound a 5-memberedheterocyclic ring selected from

wherein M is an alkali metal selected in the group consisting of lithium(Li), sodium (Na), potassium (K), rubidium (Rb) and cesium (Cs).Preferably, M is lithium (Li), sodium (Na) and potassium (K).

In a fourth embodiment of the invention, in the compounds of formula(I), -Q₁ is a cyclic nitroxide radical, as represented below

wherein

R¹ and R² are joined, as indicated by

to form together with the nitrogen atom to which they are bound a 5- to8-membered heterocyclic ring

and which may contain an additional heteroatom or heteroatomic groupselected from P(O)(OR¹¹), P(O)(R¹¹), O, S, N⁺—O⁻, NH, N(C₁₋C₆ alkyl)wherein the alkyl is straight, branched or cyclic, wherein theheterocyclic ring bears from one substituent to the maximum number ofsubstituent on the carbon atoms and optionally contains one double bond;and with the proviso that the two groups R¹ or R² together do notcontain more than one hydrogen alpha to the (N—O) group;

R¹¹ is linear or branched C₁₋₁₈ alkyl, H or an alkali metal;

X₃ is C or N with the proviso that when X₃ is N, then R⁵ is not presentin the molecule;

R³ and R⁴ are joined, through the double bond C═X₃ to form together withthe nitrogen atom to which they are bound a 5- to 8-memberedheterocyclic ring and which may contain an additional heteroatom orheteroatomic group selected from P(O)(OR¹¹), P(O)(R¹¹), O, S, N⁺—O³¹ ,NH, N(C₁₋C₆ alkyl) wherein the alkyl is straight, branched or cyclic,wherein the heterocyclic ring bears from one substituent to the maximumnumber of substituent on the carbon atoms and optionally contains onedouble bond; and with the proviso that the two groups R³ or R⁴ togetherdo not contain more than one hydrogen alpha to the (N—O) group.

In this fourth embodiment, X₁, X₂, R⁵- R¹⁰, m, n, p and q are asdescribed above. In this fourth embodiment, Q₁ is a nitroxide-containinga 5- to 8-membered heterocyclic ring, substituted at least at allpositions alpha to the (N—O) group, selected from

wherein

-   -   z is 1 or 2;    -   Y is selected from —O—, —S—, —NR with R being hydrogen,        substituted or unsubstituted linear, branched or cyclic C₁₋₆        alkyl, or —CHR—with R being hydrogen, substituted or        unsubstituted linear, branched or cyclic C₁₋₆ alkyl;    -   R¹³ and R¹⁴ are, independently, hydrogen, hydroxyl, substituted        or unsubstituted linear, branched or cyclic C₁₋₆ alkyl;    -   R¹² is, independently, hydrogen, hydroxyl, substituted or        unsubstituted linear, branched or cyclic C₁₋₆ alkyl, or    -   two geminal R¹² are joined to form together with the secondary        carbon to which they are bound a substituted or unsubstituted        cyclopentane, cyclohexane, a 5- or 6-membered heterocyclic ring        wherein at least one ring atom is oxygen;    -   R³ and R⁴ are joined, through the double bond C═X₃ to form        together with the nitrogen atom to which they are bound a 5- to        8-membered heterocyclic ring, as represented below

wherein

-   -   z is 1 or 2;    -   X₃ is C or N with the proviso that when X₃ is N, then R⁵ is not        present in the molecule;    -   R⁵ is H; (CH₂)_(n)—COOH with n being an integer from 1 to 10,        —OH, —NH₂, —(CH₂—CH₂—O)_(m)—CH₃ or (CH₂—CH₂—O)_(m)—H with m        being an integer from 1 to 10;    -   R¹⁵ and R¹⁶ are, independently, hydroxyl, substituted or        unsubstituted linear, branched or cyclic C₁₋₆ alkyl; or    -   two geminal R¹⁵ or R¹⁶ are joined to form together with the        secondary carbon to which they are bound a substituted or        unsubstituted cyclopentane, cyclohexane, a 5- or 6-membered        heterocyclic ring wherein at least one ring atom is oxygen.

In this fourth embodiment, preferably, Q₁ is

with M being an alkali metal selected in the group consisting of lithium(Li), sodium (Na), potassium (K), rubidium (Rb) and cesium (Cs).Preferably, M is lithium (Li), sodium (Na) and potassium (K),

and

-   -   R³ and R⁴ are joined and form together with the nitrogen atom to        which they are bound a 5-membered heterocyclic ring selected        from

with M being an alkali metal selected in the group consisting of lithium(Li), sodium (Na), potassium (K), rubidium (Rb) and cesium (Cs).Preferably, M is lithium (Li), sodium (Na) and potassium (K),

In all the embodiments disclosed herein, X₁ is preferably O. In all theembodiments disclosed herein, X₂ is preferably O or —NR⁹ wherein R⁹ isH; a substituted or unsubstituted, linear, branched or cyclic C₁₋₆aliphatic group; —(CH₂),—COOH with n being an integer from 1 to 10,—(CH₂—CH₂—O)_(m)—CH₃ or —(CH₂—CH₂—O)_(m)—H with m being an integer from1 to 500, preferably from 1 to 100, more preferably from 1 to 10.

In all the embodiments disclosed herein, X₂ is preferably O. In all theembodiments disclosed herein, X₂ is more preferably —NR⁹ wherein R⁹ isH; a substituted or unsubstituted, linear, branched or cyclic C₁₋₆aliphatic group; —(CH₂—CH₂—O)_(m)— CH₃ or —(CH₂—CH₂—O)_(m)—H with mbeing an integer from 1 to 10.

In all the embodiments disclosed herein, X₂ is still more preferably—NR⁹ wherein

R⁹ is H, a linear or branched C₁₋₆ alkyl, —(CH₂—CH₂—O)_(m)—CH₃ with mbeing 1 to 10.

In a fifth embodiment of the invention, the compound of formula (I) is

with X₂ as defined above;with the proviso that the compound of formula

is excluded.

The presence of “small” cycles in the compounds of formula (I) createsstrong interactions between electronic spins.

Compound 3 (referred to as AsymPol)

described herein is excluded from the definition of the compounds offormula (I). In an exemplary embodiment according to the invention, thecompound of formula (I) is compound 7:

In an exemplary embodiment according to the invention, the compound offormula (I) is compound 5:

In an exemplary embodiment according to the invention, the compound offormula (I) is compound 10 (referred to as AsymPol II):

In an exemplary embodiment according to the invention, the compound offormula (I) is compound 13:

In an exemplary embodiment according to the invention, the compound offormula (I) is compound 14: (referred to as AsymPolPOK):

Syntheses of particular compounds of formula (I) according to theinvention are described in details in the Examples.

In a general manner, syntheses of these biradicals were done by couplingof respective carboxy and amino components using peptide coupling agentsthat are commonly used during peptide synthesis (A. El-Faham and F.Albericio, Chem. Rev., 2011, 111, 6557-6602). Here, we used dicyclohexylcarbodiimide (DCC) and 1- Hydroxybenzotriazole (HOBt) as a couplingagents for the preparation of biradicals.

A general method for the synthesis of the AsymPol family of biradicalsis given below.

The compounds of the invention can be obtained according to a one-stepsynthesis as shown below:

wherein R¹⁷ and R¹⁸ are cyclic or acyclic nitroxides as defined above.The carboxy component (R¹⁷—COOH) and amino component (NH₂R¹⁸) are thestarting materials, and DCC/HOBt are reagents.

The carboxy component (R¹⁷—COOH) can be either activated in-situ or byusing its more reactive derivatives, such as acyl halides, acyl azides,acylimidazoles, anhydrides or esters. Reaction of an activated form ofthe carboxy component with the amino component (NH2R18) will form anamide as the desired coupling product.

For in-situ activation of the carboxy component (R¹⁷—COOH), couplingagents that are commonly used for peptide synthesis can be used, such ascarbonyl diimidazole (CDI), N,N′-carbonylbis(3-methylimidazolium)triflate (CBMIT), diisopropyl carbodiimide (DIC), dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3′-dimethylamino)carbodiimide.HC1 (EDCI),benzotriazol-l-yl-oxy oxytris-(dimethylamino)-phosphoniumhexafluorophosphate (BOP),benzotriazol-1-yl-oxy-tris-pyrrolidinophosphonium hexafluorophosphate(PyBop) and phosphonium-based coupling reagents, with or withouthydroxybenzotriazole (HOBt) or hydroxy-7-azabenzotriazole (HOAt) as aco-reagent. More preferably DCC/HoBt was used as a coupling agent forthe biradical synthesis.

The solvents used for the reaction may be the same or selected fromdiethylether, dimethylether, dioxane, dichloromethane, dichloroethane,acetonitrile, chloroform, dimethylformamide (DMF), dimethylsulfoxide(DMSO), tetrahydrofurane (THF) or toluene.

A tertiary amine that was used during the reaction can be triethyl amine(Et3N), N,N-diisopropylethylamine (Hunig's base or DIPEA) or4-dimethylaminopyridine (DMAP); more preferably triethyl amine.

The reaction temperature can be between 20 and 50° C., preferablybetween 20 and 30° C.

The reaction time can be between 1 and 48 h, more preferably between 6and 24 h.

The molar ratio between carboxy component (R¹⁷—COOH) and amino component(NH₂R¹⁸) can be between 1 and 1.5, in particular between 1 and 1.2.

The molar ratio between carboxy component (R¹⁷—COOH) and the reagent DCCcan be between 1 and 1.5, in particular between 1 and 1.2.

The molar ratio between carboxy component (R¹⁷—COOH) and the reagentHOBt can be between 1 and 5, in particular between 2 and 3.

The molar ratio between carboxy component (R¹⁷—COOH) and the Et₃N can bebetween 1 and 5, in particular between 2 and 4wherein R¹⁷ and R¹⁸ arecyclic or acyclic nitroxides as defined above. The carboxy component(R¹⁷—COOH) and amino component (NH₂R¹⁸) are the starting materials, andDCC/HOBt are reagents.

The desired product of the coupling reaction can be isolated andpurified by conventional separation/purification methods used in thesynthetic organic chemistry, such as filtration, extraction, washing,drying, concentration, recrystallization, various chromatographictechniques or the like.

The compounds of the invention are stable and are soluble either inaqueous or organic solvent. Another object of the present inventionrelates to the use of at least one compound of formula (I) according tothe invention or a compound of formula

as a polarizing agent.

Another object of the invention relates to the use of at least onecompound of formula

(I) according to the invention or a compound of formula

as a polarizing agent in the techniques of structural biology, NuclearMagnetic Resonance (NMR) of solids or applied to liquid samples,particle physics, and medical imaging.

In particular, the compounds according to the invention or a compound offormula

may be used as DNP agents for polarizing an NMR-active isotope of anucleus in Nuclear Magnetic Resonance (NMR) spectroscopy. The term NMRspectroscopy, as used herein, encompasses Solid State NMR (SS-NMR)spectroscopy, liquid state NMR spectroscopy and Magnetic ResonanceImaging (MRI), in all of which the biradical compounds of the inventionmay be used as DNP agents.

A nucleus having an NMR-active spin may be, for example: ¹H, ²H, ⁶Li,⁷Li, ¹⁰B, ¹¹B, ¹³C, ¹⁴N, ¹⁵N, ¹⁷O, ¹⁹F, ²³Na, ²⁵Mg, ²⁷Al, ²⁹Si, ³¹P,³³S, ³⁵Cl, ³⁷Cl, ³⁹K, ⁴¹K, ⁴³Ca, ⁴⁷Ti, ⁴⁹Ti, ⁵⁰Vm ⁵¹V, ⁵³Cr, ⁷⁷Se, ⁸⁹Y,¹¹⁷Sn, ¹¹⁹Sn and ¹⁹⁹Hg.

A further object of the invention relates to a method for polarizing acompound in a sample for Dynamic Nuclear Polarization comprisingcontacting said sample with at least one compound of formula (I)according to the invention.

The method for polarizing a compound in a sample for Dynamic NuclearPolarization according to the invention comprises the steps of:

-   -   a) providing said at least one compound of formula (I) or a        compound of formula

as polarizing agent that enables an optimal nuclear polarization of thesample in a magnetic field;

-   -   b) irradiating said sample comprising the compound of        formula (I) or a compound of formula

with at least one radiation that causes electron spin flip, to enhancethe performance of NMR detection or MRI performance; and

-   -   c) optionally dissolving the sample and obtaining a        hyperpolarized sample.

Said method may further comprise observing the NMR or MRI of thehyperpolarized compound of formula (I).

The irradiation is preferably a microwave irradiation. The frequencyrange of the microwave irradiation by which the polarization istransferred to an NMR-active nucleus is usually from 5 to 800 GHz.

The term “sample”, as used herein, refers to a chemical or a biologicalentity, such as a solid inorganic, organic or metallo-organic materialhaving a crystal lattice or an amorphous solid structure (e.g. zeolites,nanoparticles, mesoporous and porous materials, glasses, Metal OrganicFrameworks (MOF), a molecular chemical or biochemical compound includingpolymeric compounds and macromolecular compounds (e.g. proteins,enzymes, DNA/RNA and a biological entity (e.g. a whole cell, a leaf, avirus particle, tissue or bone components or a whole body, having one ormore NMR-active spins to be investigated by NMR spectroscopy. Thechemical or a biological entity may be isolated or in its naturalenvironment. The sample may be dissolved in aqueous medium, an organicsolvent or a mixture of organic or aqueous/organic solvents. The samplemay be without a solvent.

The investigation by NMR spectroscopy may be structure determination,monitoring of reaction kinetics, flow imaging, etc.

The polarizing agent may be dissolved in the solvent of the sample orcan be chemically bound to the sample and be introduced without asolvent.

The polarizing agent may be present in a solid state, with or without asolvent or mixture of solvents (e.g. frozen solvent(s)), during thepolarization time.

The polarizing agent may be present in a liquid state or in a liquidsolution during the polarization time.

A compound of formula (I) according to the invention or a compound offormula

when used as a polarizing agent, is used at a concentration of 0.01 to200 mM.

In solid state NMR experiments, the temperature of a sample includingpolarizing agent is in the range of 1 to 300 K.

The invention will be further illustrated by the following figures andexamples. These examples and figures should not in any way beinterpreted as limiting the scope of the present invention.

FIGURES

FIG. 1 represents the EPR spectrum of Compound 3, in CH₂Cl₂ (1 mM) at25° C.

FIG. 2 represents the ¹H-NMR of Compound 3 in CDCl₃.

FIG. 3 represents the HPLC chromatogram of Compound 3.

FIG. 4 represents the EPR spectrum of Compound 5, in CH₂Cl₂ (1 mM) at25° C.

FIG. 5 represents the ¹H-NMR of Compound 5 in CDCI₃.

FIG. 6 represents the HPLC chromatogram of Compound 5.

FIG. 7 represents the EPR spectrum of Compound 7, in CH₂Cl₂ (1 mM) at25° C.

FIG. 8 represents the ¹H-NMR of Compound 7 in CDC₃.

FIG. 9 represents the HPLC chromatogram of Compound 7.

FIG. 10 represents the ¹H-NMR of Compound 9 in D₂O.

FIG. 11 represents the EPR spectrum of Compound 10, in water (1 mM) at25° C.

FIG. 12 represents the ¹H-NMR of Compound 10, in D₂O.

FIG. 13 represents the HPLC chromatogram of Compound 10.

FIG. 14 represents the ¹H-NMR of Compound 13 in CDCl₃.

FIG. 15 represents the EPR spectrum of Compound 14, in water (1 mM) at25° C.

FIG. 16 represents the ¹H-NMR of Compound 14 in in D₂O.

FIG. 17 represents the HPLC chromatogram of Compound 14.

EXAMPLES 1/Synthesis of the AsymPol Biradical Family

Chemicals were purchased primarily from the Sigma-Aldrich ChemicalCompany and Acros and were used without further purification.Dichloromethane, acetonitrile and pyridine were freshly distilled overcalcium hydride before use; triethylamine was purchased anhydrous andstored over potassium hydroxide pellets. Thin layer chromatography (TLC)was performed on glass backed TLC plates with extra hard layer(Kieselgel 60 F₂₅₄, 250 μm, Silicycle) and compounds were visualized byUV light. Silica gel (230-400 mesh, 60 Å) was purchased from Silicycle,and used for flash chromatography. ¹H and ¹³C NMR spectra were recordedat the frequencies stated, using deuterated solvents as internalstandards. 400 MHz spectra were recorded on a Bruker Advance 400spectrometer. Residual proton signals from the deuterated solvents wereused as references [D₂O (4.81 ppm), <d₆-DMSO (2.50 ppm), chloroform(7.26 ppm), <d₄-MeOH (4.84 and 3.31 ppm)] for ¹H spectra. The residual¹³C signals from the deuterated solvents being used as references[J6-DMSO (39.7 ppm), chloroform (77.0 ppm), <d₄-MeOH (49.05 ppm)] for¹³C spectra. All coupling constants were measured in Hertz. All moisturesensitive reactions were carried out in oven-dried glassware usingnitrogen or argon from standard BOC industrial cylinders, dried throughan activated silica column. Molecular mass of the new organic compoundswas determined by HR-ESI/ESI-MS (Bruker, MicroTof-Q). Purity of 3, 5, 10and 14 were analysed on GL Sciences Inertsustain C18 4.6×150 mmanalytical column with UV detection at 254 nm on Beckman Coulter GoldHPLC system. Analytical HPLC run (Flow rate=1 mL/min): Solvent A, 0.1%TFA in water; Solvent B, MeOH; 8 min linear gradient from 0% to 100% B,2 min linear gradient from 100% to 0% B (initial conditions; 100% A).

Compound 3: To a solution of 2 (K. Oyaizu, T. Kawamoto, T. Suga and H.Nishide, Macromolecules, 2010, 43, 10382-10389) (0.032 g, 0.175 mmol) inCH₂Cl₂ (4 mL) was added DCC (0.039 g, 0.192 mmol), HOBt (0.053 g, 0.35mmol) and triethyl amine (0.073 mL, 0.525 mmol) under an inertatmosphere of argon. After stirring for 15 minutes, 1 (G. M. Rosen, J.Med. Chem., 1974, 17, 358-360) (0.03 g, 0.175 mmol) was added. Theresulting solution was stirred for 12 hours at 25° C., diluted withCH₂Cl₂ (10 mL) and washed successively with sat. aqueous solution ofNaHCO₃ (10 mL) and brine (10 mL). The organic layer was concentrated invacuo and the crude product was purified by flash column chromatography(silica) using a gradient elution (EtOAc: petroleum ether; 0: 100 to30:70) to give 3 (0.049 g, 83% yield) as a yellow solid.

TLC (Silica gel, 10% MeOH in CH₂Cl₂), R_(f)(1)=0.2, R_(f)(2)=0.8,R_(f)(3)=0.9, PMA active.

(Silica gel, 2.5% MeOH in CH₂Cl₂), R_(f)(3)=0.3

¹H-NMR (CDCl₃): Compound 3 is a nitroxide biradical and hence, showsbroadening of peaks. Therefore, integration of the NMR peaks in NMRspectra was not performed.

LCMS: calculated for C₁₈H₃₁N₃O₃: 337.2365, found 339.2520 (M+2H) ²⁺.

Compound 5: To a solution of 2 (K. Oyaizu, T. Kawamoto, T. Suga and H.Nishide, Macromolecules, 2010, 43, 10382-10389) (0.03 g, 0.162 mmol) inCH₂Cl₂ (4 mL) was added DCC (0.037 g, 0.18 mmol), HOBt (0.049 g, 0.32mmol) and triethyl amine (0.07 mL, 0.495 mmol) under an inert atmosphereof argon. After stirring for 15 minutes, 4 (G. M. Rosen, J. Med. Chem.,1974, 17, 358-360) (0.03 g, 0.162 mmol) was added. The resultingsolution was stirred for 12 hours at 25° C., diluted with CH₂Cl₂ (10 mL)and washed successively with sat. aqueous solution of NaHCO₃ (10 mL) andbrine (10 mL). The organic layer was concentrated in vacuo and the crudeproduct which was purified by flash column chromatography (silica) usinga gradient elution (EtOAc: petroleum ether; 0:100 to 35:65) to give 5(0.051 g, 91% yield) as a yellow solid.

TLC (Silica gel, 40% EtOAc in pet ether), R_(f)(2)=0.3, R_(f)(5)=0.2,PMA active.

¹H-NMR (CDCl₃): Compound 5 is a nitroxide biradical and hence, showsbroadening of peaks. Therefore, integration of the NMR peaks in NMRspectra was not performed.

LCMS: calculated for C₁₉H₃₃N₃O₃: 351.2522, found 353.2673 (M+2H)²⁺.

Compound 7: To a solution of 2 (K. Oyaizu, T. Kawamoto, T. Suga and H.Nishide, Macromolecules, 2010, 43, 10382-10389) (0.06 g, 0.326 mmol) inTHF (6 mL) was added DCC (0.087 g, 0.423 mmol), HOBt (0.057 g, 0.423mmol) and DMAP (0.23 g, 0.195 mmol) under an inert atmosphere of argon.After stirring for 15 minutes, 6 (0.068 g, 0.390 mmol) was added. Theresulting solution was stirred for 12 hours at 25° C. Solvent wasremoved under vacuo. The residue was diluted with CH₂Cl₂ (10 mL) andwashed successively with sat. aqueous solution of NaHCO₃ (10 mL) andbrine (10 mL). The organic layer was concentrated in vacuo and the crudeproduct which was purified by flash column chromatography (silica) usinga gradient elution (EtOAc: petroleum ether; 0: 100 to 20:80) to give 7(0.048 g, 43% yield) as a yellow solid.

TLC (Silica gel, 20% EtOAc in pet ether), R_(f)(2)=0.3, R_(f)(7)=0.2,PMA active.

¹H-NMR (CDCl₃): Compound 7 is a nitroxide biradical and hence, showsbroadening of peaks. Therefore, integration of the NMR peaks in NMRspectra was not performed.

HRMS: calculated for C₁₈H₃₀N₂O₄: 338.4480, found 361.2096 (M+Na)⁺.

Compound 9: To a solution of 2 (K. Oyaizu, T. Kawamoto, T. Suga and H.Nishide, Macromolecules, 2010, 43, 10382-10389) (0.005 g, 0.029 mmol) inCH₂Cl₂ (3 mL) was added DCC (0.006 g, 0.32 mmol), HOBt (0.009 g, 0.059mmol) and triethylamine (0.013 mL, 0.088 mmol) under an inert atmosphereof argon. After stirring for 15 minutes, 8 (A. P. Jagtap, M. A. Geiger,D. Stoppler, M. Orwick-Rydmark, H. Oschkinat and S. T. Sigurdsson, Chem.Commun., 2016, 52, 7020-7023) (0.015 g, 0.029 mmol) was added. Theresulting solution was stirred for 12 hours at 25° C. The reactionmixture was diluted with CH₂Cl₂ (10 mL) and washed successively withsat. aqueous solution of NaHCO₃ (10 mL) and brine (10 mL). The organiclayer was concentrated in vacuo and the crude product which was purifiedby flash column chromatography (silica) using a gradient elution (EtOAc:petroleum ether; 0:100 to 30:70) to give 9 (0.011 g, 58% yield) as ayellow solid.

TLC (Silica gel, 3% Me0H in CH₂Cl₂), R_(f)(2)=0.3, R_(f)(9)=0.6, PMAactive.

¹H-NMR (CDCl₃): Compound 9 is a nitroxide biradical and hence, showsbroadening of peaks. Therefore, integration of the NMR peaks in NMRspectra was not performed.

HRMS: calculated for C₃₆H₆₇N₃O₅Si₂: 677.4619, found 700.4505 (M+Na)⁺.

Compound 10: TBAF (0.8 mL, 0.778 mmol, 1 M in THF) was added to asolution of 9 (0.088 g, 0.130 mmol) in anhydrous THF (4 mL). Theresulting solution was heated at 60° C. for 12 hours, cooled down andthe solvent removed in vacuo. The residue was dissolved in MeOH (4 mL)and DOWEX (0.50 g) and CaCO₃ (0.165 g) were added. The resultingsuspension was stirred at for 12 hours at 27° C. The reaction mixturewas filtered through a bed of celite, the filtrate concentrated undervacuo and the crude product was purified by flash column chromatography(silica) using a gradient elution (MeOH: CH₂Cl₂; 0:100 to 10:90) to give10 (0.034 g, 49% yield) as a yellow solid.

TLC (Silica gel, 10% MeOH in CH₂Cl₂), R_(f)(12)=1, R_(f)(10)=0.1, PMAactive.

¹H-NMR (D₂O): Compound 10 is a nitroxide biradical and hence, showsbroadening of peaks. Therefore, integration of the NMR peaks in NMRspectra was not performed.

HRMS: calculated for C₂₄H₃₉N₃O₅: 449.2890, found 472.2774 (M+Na)⁺.

Compound 13: To a solution of 10 (0.045 g, 0.10 mmol) in CH₃CN (4 mL)was added 11 (0.058 g, 0.3 mmol) and 12 (0.053 g, 0.35 mmol) under aninert atmosphere of argon. After stirring for 2 hours at 27° C., tBuOOH(0.384 mL, 3.2 mmol, 75% in water) was added. The resulting solution wasstirred for 30 minutes at 25° C. The solvent was removed under vacuo,the residue obtained was diluted with CH₂Cl₂ (10 mL) and washedsuccessively with sat. aqueous solution of NaHCO₃ (10 mL) and brine (10mL). The organic layer was concentrated in vacuo and the crude productwas purified by flash column chromatography (silica) using a gradientelution (MeOH: CH₂C12; 0:100 to 3:97) to give 13 (0.028 g, 35% yield) asa yellow solid.

TLC (Silica gel, 10% MeOH in CH₂Cl₂), R_(f)(10)=0.4, R_(f)(13)=0.6, PMAactive.

¹H-NMR (CDCl₃): Compound 13 is a nitroxide biradical and hence, showsbroadening of peaks. Therefore, integration of the NMR peaks in NMRspectra was not performed.

³¹P-NMR (CDCl₃): -3.74, -4.35

HRMS: calculated for C₃₆H₅₃N₇O₁₁P₂: 821.3278, found 844.3172 (M+Na).

Compound 14: To a solution of 13 (0.045 g, 0.0739 mmol) in H₂O (2 mL)was added triethylamine (0.3 mL, 2.143 mmol). The resulting solution wasstirred for 12 hours at 60° C. The solvent was removed in vacuo. Theresidue was diluted with H₂O (2 mL) and KOH (0.018 g, 0.325 mmol) wasadded. The resulting solution was stirred for 12 hours at 60 ° C. Thesolvent was removed in vacuo to give 14 (0.038 g, 68%>yield) as a yellowsolid.

TLC (Silica gel, 10% MeOH in CH₂Cl₂), R_(f)(13)=0.6, R_(f)(14)=O,PMAactive.

¹H-NMR (D20): Compound 14 is a nitroxide biradical and hence, showsbroadening of peaks. Therefore, integration of the NMR peaks in NMRspectra was not performed.

³¹P-NMR (D20): −3.23

HRMS: calculated for C₂₄H₄₁N₃O₁₁P₂: 609.2216, found 630.1948 (M+Na−2H)

2/ Sensitivity of the Compounds According to the Invention Under DNPConditions

The table below compares the sensitivity gain (ϵ_(B)/√{square root over(T_(B) ^(T))}⁺ ) obtained with compound according to the invention,AsymPol, AsymPolPOK and AMUPol which is currently considered as one ofthe best performing polarizing agent for MAS-DNP experiments.

TABLE 1 Experimental parameters that characterize the DNP performance ofAsymPol and AsymPolPOK, with a comparison to AMUPol. Buildup Time DNPsensitivity DNP gain T_(B) ^(ε)on/off ε_(B) · T_(B(MAS)) _(−1/2) ε_(B)_((MAS)) ⁻ Static MAS Static MAS 9.4 T AMUPol^([a]) 27 s ^(−1/2) 43 16.3s 2.5 s 28 151 AsymPol^([b]) 39 s ^(−1/2) 30 1.0 s 0.6 s 11 32AsymPolPOK^([a]) 68 s ^(−1/2) 83 3.5 s 1.5 s 25 105 ^([a])10 mMbiradical in d₈-glycerol/D₂O/H₂0 (6:3:1; v:v) with 20 mM ¹³C-urea, 10kHz MAS rate, at 105K and 9.4 T. ^([b])same as [a] but 10 mM biradicalin d₆- DMSO/D_(2O)/H₂O(8:1:1; v:v). ^([c])same as [a] but at −130K and 8kHz MAS rate using a 3.2 mm rotor.

It appears clearly from these results that the use of AsymPol biradicalsprovides a significant increase in DNP sensitivity compared to AMUPol.These results also illustrate the limits of relying solely on Bon Off toevaluate polarizing agent's efficiency.

3/ Characteristics of the Compounds According to the Invention

Table 2 compares certain characteristics of the known polarizing agents,i.e. TOTAPOL, AMUPol and TEKPol.

TABLE 2 AMUPol TEKPol (« b Turea » (« bTbK » Characteristics AsymPolTOTAPOL family) family) Spin interaction intense moderate moderate tomoderate intense Relative rigid flexible rigid very rigid orientationsof the g tensor Amplification important moderate important to importantfactor ε_(on/off) (for very to very 10 mM biradicals) importantimportant Depolarization weak moderate strong moderate strength (almostto strong absent Polarization speed very sast slow fast moderateSensitivity gain very high moderate moderate to moderate to importantimportant Efficiency at high good weak weak to weak MAS frequencymoderate (>20 kHz)

The table above illustrates the reason for which the polarizing agentsproposed up now offer modest performances (especially at high spinningfrequencies).

1. A compound of formula (I)

wherein X₁ is O, C(R⁶R⁷), NR⁸, S; X₂ is O, SO₂, or —NR⁹, CH₂; X₃ is C orN with the proviso that when X₃ is N, then R⁵ is not present in themolecule; R⁵, R⁶, R⁷, R⁸ and R⁹ are, independently, H; a substituted orunsubstituted, linear, branched or cyclic C₁₋₆ aliphatic group;—(CH₂)_(n)—COOH with n being an integer from 1 to 10, —OH, —NH₂,—N₃,—C≡CH, P(O)(OH)₂, P(O)(OR¹¹)₂, P(O)R¹¹ ₂, SSO₂Me, —(CH₂—CH₂—O)₃—CH₃ or—(CH₂—CH₂—O)_(m)—H with m being an integer from 1 to 500, preferablyfrom 1 to 100, more preferably from 1 to 10, or

 with p being an integer from 0 to 7 and q an integer from 1 to 500; Q₁is a cyclic or acyclic nitroxide radical, as represented below

wherein R¹ and R² are, independently, a substituted or unsubstituted,linear, branched or cyclic C₁₋₆ aliphatic group; a substituted orunsubstituted linear, branched or cyclic hetero-aliphatic groupcomprising 5 carbon atoms and one heteroatom or heteroatomic group,respectively, selected from O,S, —NR¹⁰—, P(O)(OR¹¹)₂ and P(O)(R¹¹)₂;substituted or unsubstituted aryl; substituted or unsubstitutedheteroaryl, or substituted or unsubstituted 2,2,7,7-tetramethylisoindolinoxyl; substituted or unsubstituted 2,2,7,7-tetraethylisoindolinoxyl; or R¹ and R² are joined, as indicated by

 to form together with the nitrogen atom to which they are bound a 5- to8-membered heterocyclic ring and which may contain an additionalheteroatom or heteroatomic group selected from P(O)(OR¹¹)₂, P(O)(R¹¹)₂,O, S, N⁺−O⁻, NH, N(C₁-C₆ alkyl) wherein the alkyl is straight, branchedor cyclic, wherein the heterocyclic ring bears from one substituent tothe maximum number of substituent on the carbon atoms and optionallycontains one double bond; and with the proviso that the two groups R¹ orR² together do not contain more than one hydrogen alpha to the (N—O.)group; R¹⁰ is hydrogen, hydroxyl; substituted or unsubstituted linear,branched or cyclic C₁₋₆ alkyl; C₁₋₆ alkylcarbonyl; substituted orunsubstituted aryl sulfinyl; or substituted or unsubstituted arylsulfonyl; R¹¹ is linear or branched C₁₋₁₈ alkyl, H or an alkali metal;and wherein the point of attachment of the X₂ atom by a single bond, asindicated by

 is to a primary or secondary non-olefinic or aromatic carbon atom ofeither R¹ or R², or to a carbon atom of the 5- to 8-memberedheterocyclic ring formed by the joining of R¹ and R²; R³ or R⁴ is linkedto the double bond C═X₃ in the compound of formula (I) as representedabove, wherein R³ and R⁴ are, independently, a substituted orunsubstituted, linear, branched or cyclic C₁₋₆ aliphatic group; asubstituted or unsubstituted linear, branched or cyclic hetero-aliphaticgroup comprising 5 carbon atoms and one heteroatom or heteroatomicgroup, respectively, selected from O,S, —NR¹⁰—, P(O)(OR¹¹)₂ andP(O)(R¹¹)₂; substituted or unsubstituted aryl; substituted orunsubstituted heteroaryl, or substituted or unsubstituted 2,2,7,7-tetramethyl isoindolinoxyl; substituted or unsubstituted2,2,7,7-tetraethyl isoindolinoxyl; or R³ and R⁴ are joined, through thedouble bond C═X₃ to form together with the nitrogen atom to which theyare bound a 5- to 8-membered heterocyclic ring and which may contain anadditional heteroatom or heteroatomic group selected from P(O)(OR¹¹)₂,P(O)(R¹¹)₂, O, S, N^(±)—O⁻, NH, N(C₁-C₆ alkyl) wherein the alkyl isstraight, branched or cyclic, wherein the heterocyclic ring bears fromone substituent to the maximum number of substituent on the carbon atomsand optionally contains one double bond; and with the proviso that thetwo groups R³ or R⁴ together do not contain more than one hydrogen alphato the (N—O.) group; with the proviso that the compound of formula

is excluded.
 2. The compound according to claim 1, wherein Q₁ is acyclic nitroxide radical, as represented below

wherein R¹ and R² are joined, as indicated by

 to form together with the nitrogen atom to which they are bound a 5- to8-membered heterocyclic ring and which may contain an additionalheteroatom or heteroatomic group selected from P(O)(OR¹¹)₂,P(O)(R¹¹)₂,O, S, N^(±)−O⁻, NH, N(C₁-C₆ alkyl) wherein the alkyl isstraight, branched or cyclic, wherein the heterocyclic ring bears fromone substituent to the maximum number of substituent on the carbon atomsand optionally contains one double bond; and with the proviso that thetwo groups R¹ or R² together do not contain more than one hydrogen alphato the (N—O.) group; and R¹¹ is linear or branched C₁₋₁₈ alkyl, H or analkali metal.
 3. The compound according to claim 1, wherein Q₁ is,

with M being an alkali metal selected in the group consisting of lithium(Li), sodium (Na), potassium (K), rubidium (Rb) and cesium (Cs),


4. The compound of formula (I) according to claim 1, wherein Q₁ is anacyclic nitroxide radical, as represented below

wherein R¹ and R² are, independently, a substituted or unsubstituted,linear, branched or cyclic C₁₋₆ aliphatic group; a substituted orunsubstituted linear, branched or cyclic hetero-aliphatic groupcomprising 5 carbon atoms and one heteroatom or heteroatomic group,respectively, selected from O,S, —NR¹⁰—, P(O)(OR¹¹) and P(O)(R¹¹);substituted or unsubstituted aryl; substituted or unsubstitutedheteroaryl; or substituted or unsubstituted 2,2,7,7-tetraethylisoindolinoxyl; R¹⁰ is hydrogen, hydroxyl; substituted or unsubstitutedlinear, branched or cyclic C₁₋₆ alkyl; C₁₋₆ alkyl carbonyl; substitutedor unsubstituted aryl sulfinyl; or substituted or unsubstituted arylsulfonyl; R¹¹ is linear or branched C₁₋₁₈ alkyl, H or an alkali metal;with the proviso that the two groups R¹ or R² together do not containmore than one hydrogen alpha to the (N—O.) group.
 5. The compound offormula (I) according to claim 1, wherein R³ and R⁴ are joined, throughthe double bond C═X₃ to form together with the nitrogen atom to whichthey are bound a 5- to 8-membered heterocyclic ring and which maycontain an additional heteroatom or heteroatomic group selected fromP(O)(OR¹¹)₂, P(O)(R¹¹)₂, O, S, N±—O⁻, NH, N(C₁-C₆ alkyl) wherein thealkyl is straight, branched or cyclic, wherein the heterocyclic ringbears from one substituent to the maximum number of substituent on thecarbon atoms and optionally contains one double bond; with X₃ as definedin claim 1 and with the proviso that the two groups R³ or R⁴ together donot contain more than one hydrogen alpha to the (N—O.) group.
 6. Thecompound of formula (I) according to claim 1, wherein R³ and R⁴ arejoined and form together with the nitrogen atom to which they are bounda 5-membered heterocyclic ring selected from

wherein M is an alkali metal selected in the group consisting of lithium(Li), sodium (Na), potassium (K), rubidium (Rb) and cesium (Cs).Preferably, M is lithium (Li), sodium (Na) or potassium (K).
 7. Thecompound of formula (I) according to claim 1, wherein X₂ is O or —NR⁹wherein R⁹ is H; a substituted or unsubstituted, linear, branched orcyclic C₁₋₆ alkyl group; —(CH₂)_(n)—COOH with n being an integer from 1to 10, —(CH₂—CH₂—O).—CH₃ or —(CH₂—CH₂—O)_(m)—H with m being an integerfrom 1 to 500, preferably from 1 to 100, more preferably from 1 to 10.8. The compound of formula (I) according to claim 1, wherein X₂ is O or—NR⁹ wherein R⁹ is H, a linear or branched C₁₋₆ alkyl,—(CH₂—CH₂—O)_(m)—CH₃ with m being 1 to
 10. 9. The compound of formula(I) according to claim 1, wherein it is represented as below

with X₂ as defined in any one of the preceding claims; with the provisothat the compound of formula

is excluded.
 10. The compound of formula (I) according to claim 1,wherein it is represented as below:


11. The compound of formula (I) according to claim 1, wherein it isrepresented as below:


12. Use of at least one compound of formula (I) according to claim 1 asa polarizing agent.
 13. The use of at least one compound of formula (I)according to claim 1 in the techniques of structural biology, NuclearMagnetic Resonance (NMR) of solids or applied to liquid samples,particle physics, and medical imaging.
 14. A method for polarizing acompound in a sample for Dynamic Nuclear Polarization comprisingcontacting said sample with at least one compound of formula (I)according to claim
 1. 15. The method for polarizing a compound in asample for Dynamic Nuclear Polarization, wherein said method comprisesthe steps of: a) providing at least one compound of formula (I)according to claim 1 as polarizing agent that enables an optimal nuclearpolarization of a sample in a magnetic field; b) irradiating said samplecomprising the compound of formula (I) with at least one radiation thatcauses electron spin flip, to enhance the performance of NMR detectionor MRI performance; and c) optionally dissolving the sample andobtaining a hyperpolarized sample.